Biomarkers are urgently needed for the early and accurate diagnosis of Alzheimer's disease (AD). SurroMed, Inc. is using high throughput, metabolomic and proteomic approaches to differentially quantify hundreds to thousands of molecules in human cerebrospinal fluid (CSF), serum, and urine to identify biomarkers of AD. The low molecular peptide and metabolite fraction (the metabolome) contains a rich diversity of possible biomarkers of AD, yet it has never been systematically examined. We propose to develop methods to fractionate the metabolome of CSF, serum, and urine and identify peptides and metabolites using micro-scale liquid chromatography and mass spectrometry. This discovery-based approach will be complemented with a hypothesis-based approach in which we examine molecules implicated in AD pathophysiology using immuno-affinity capture and/or MS. Phase I will therefore establish a metabolomic platform for detection and identification of AD biomarkers at an unprecedented scale and with high throughput. This technology will be applied in a subsequent Phase II application aimed at discovering biomarkers for early stages of AD in a clinical study with mild cognitively impaired and AD patients.